Phase I: High throughput fabrication of fractal cell culture substrates

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Currently, there are no cell culture techniques that enable high fidelity cultivation of the key kidney cells: podocytes. Monolayer culture fails to recapitulate their branching phenotype. We have developed a completely new approach: by faithfully recreating fractal topographical cues of the native kidney glomerulus on cell culture substrates, we microengineered an environment where podocytes exhibit their branching morphology and gene expression similar to the podocytes in vivo. Such improved cell culture was superior in new compound testing and coronavirus infection studies compared to the conventional monolayers. The main goal of this proposal is to develop a high throughput manufacturing process for production of commercial well plates with fractal topographical cues, from the current manual process that takes >20hr to produce one 2x2cm2 substrate, to a process that takes 15min per plate. In Aim 1, we will scale-up production by hot embossing and injection molding to create commercial 24- and 96-well plates. In Aim 2, we will develop sterilization protocol for these plates and in Aim 3, we will validate their biocompatibility. To reach the desired production throughput and create non-absorbent devices, we need to overcome the main engineering challenge of scaling from polydimethylsiloxane lab prototype inserts to non-absorbent thermoplastic elastomer laminated on glass. Our main customers will be pharmaceutical companies, e.g. Regeneron that filled an order form, and academic laboratories who have a need for high fidelity culture of podocytes. Our collaborators, Drs. Quaggin, Barua and Rogers, are leading kidney researchers, eager to test the new plates for podocyte cultivation in their laboratories. The new manufacturing process can also be used to fabricate fractal plates for other cells e.g. adipocytes and lung cells. Our preferred model is licensing the plates to an existing established company that manufactures and distributes innovative tools for life science research, such as companies that provided support letters-Stem Cell Technologies and uFluidix. The plates will also enable discovery of new compounds for kidney disease, through a new start-up that our team members will work on.